Cell Signaling Technology

Product Pathways - MAPK Signaling

PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit #9100

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 200 µl W IP IHC-P IF-IC F H M R Hm Mk Mi Dm Z B Dg Pg Sc (C) (Ce) 44, 42 Rabbit IgG
p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 200 µl W IP IHC-P IF-IC F H M R Hm Mk Mi Dm Z B Dg Pg Ce (C) 42, 44 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat
p44/42 MAPK (Erk1/2) Control Proteins #9103 100 µl 42
Anti-biotin, HRP-linked Antibody #7075 250 µl Goat
Biotinylated Protein Ladder Detection Pack #7727 250 µl
20X LumiGLO® Reagent and 20X Peroxide #7003 10 ml

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  C=Chicken  Mi=Mink  Dm=D. melanogaster  Z=Zebrafish  B=Bovine  Dg=Dog  Pg=Pig  Sc=S. cerevisiae  Ce=C. elegans
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) Rabbit mAb #4370 detects endogenous levels of p42 and p44 MAP Kinase (Erk1 and Erk2) when dually phosphorylated at Thr202 and Tyr204 of Erk1 (Thr185 and Tyr187 of Erk2), and singly phosphorylated at Thr202. This antibody does not cross-react with the corresponding phosphorylated residues of either JNK/SAPK or p38 MAP kinases. p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 detects endogenous levels of total p44/42 MAP kinase (Erk1/Erk2) protein. The antibody does not cross-react with either JNK/SAPK or p38 MAP kinases.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293, NIH/3T3 and C6 cells, treated with λ phosphatase or TPA as indicated, using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) Rabbit mAb #4370 (upper), or p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) Rabbit mAb #4370.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 in the presence of control peptide (left) or #1240 p44/42 MAPK (Erk1/2) Blocking Peptide (#4695 Specific) (right).


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, U0126-treated (blue) or PMA-treated (green), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) Rabbit mAb #4370.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, U0126-treated (blue) or PMA-treated (green), using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 compared to a nonspecific negative control antibody (red).

IF-IC

IF-IC

Confocal immunofluorescent analysis of C2C12 cells, U0126-treated (left, #9903 10 μM for 1 h) or TPA-treated (right, #9905 200 nM for 15 min), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) Rabbit mAb #4370 (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).


IF-IC

IF-IC

Confocal immunofluorescent analysis of NIH/3T3 cells either U0126-treated (left) or PDGF-treated (right) and labeled with p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

Description

The PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit provides reagents and protocols for the rapid analysis of the phosphorylation status of p44 and p42 (Erk1 and Erk2). Bacterially expressed, kinase inactive p42 MAP kinase (Erk2) protein serves as a negative control. The same protein phosphorylated by MEK2 serves as a positive control.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr202/Tyr204 of human p44 MAP kinase or a synthetic peptide corresponding to residues near the C-terminus of rat p44 MAP kinase.

Background

Mitogen-activated protein kinases (MAPKs) are a widely conserved family of serine/threonine protein kinases involved in many cellular programs, such as cell proliferation, differentiation, motility, and death. The p44/42 MAPK (Erk1/2) signaling pathway can be activated in response to a diverse range of extracellular stimuli including mitogens, growth factors, and cytokines (1-3), and research investigators consider it an important target in the diagnosis and treatment of cancer (4). Upon stimulation, a sequential three-part protein kinase cascade is initiated, consisting of a MAP kinase kinase kinase (MAPKKK or MAP3K), a MAP kinase kinase (MAPKK or MAP2K), and a MAP kinase (MAPK). Multiple p44/42 MAP3Ks have been identified, including members of the Raf family, as well as Mos and Tpl2/COT. MEK1 and MEK2 are the primary MAPKKs in this pathway (5,6). MEK1 and MEK2 activate p44 and p42 through phosphorylation of activation loop residues Thr202/Tyr204 and Thr185/Tyr187, respectively. Several downstream targets of p44/42 have been identified, including p90RSK (7) and the transcription factor Elk-1 (8,9). p44/42 are negatively regulated by a family of dual-specificity (Thr/Tyr) MAPK phosphatases, known as DUSPs or MKPs (10), along with MEK inhibitors, such as U0126 and PD98059.

  1. Roux, P.P. and Blenis, J. (2004) Microbiol Mol Biol Rev 68, 320-44.
  2. Baccarini, M. (2005) FEBS Lett 579, 3271-7.
  3. Meloche, S. and Pouysségur, J. (2007) Oncogene 26, 3227-39.
  4. Roberts, P.J. and Der, C.J. (2007) Oncogene 26, 3291-310.
  5. Rubinfeld, H. and Seger, R. (2005) Mol Biotechnol 31, 151-74.
  6. Murphy, L.O. and Blenis, J. (2006) Trends Biochem Sci 31, 268-75.
  7. Dalby, K.N. et al. (1998) J Biol Chem 273, 1496-505.
  8. Marais, R. et al. (1993) Cell 73, 381-93.
  9. Kortenjann, M. et al. (1994) Mol Cell Biol 14, 4815-24.
  10. Owens, D.M. and Keyse, S.M. (2007) Oncogene 26, 3203-13.

Application References

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Protocols

* Product-specific protocol.

Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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